Analysis of poly(ADP-ribose) function in the cytoplasmic stress response

细胞质应激反应中聚（ADP-核糖）功能的分析

• 批准号: 8707478
• 负责人: PAUL CHANG
• 金额: $ 30.5万
• 依托单位: MASSACHUSETTS INSTITUTE OF TECHNOLOGY
• 依托单位国家: 美国
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-09-30 至 2016-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8707478
• 关键词: Adenosine Diphosphate Ribose; Apoptosis; Binding; Binding Proteins; Biochemical; Biological Assay; Biological Process; Cell Cycle Progression; Cell division; Cell physiology; Cells; Cellular Stress; Cellular biology; Characteristics; Clinical Treatment; Cytoplasm; Cytoplasmic Granules; DNA Damage; DNA Repair; Disease; Enzymes; Family; Goals; Heat-Shock Response; Immune response; Ischemia; Life; Malignant Neoplasms; Malignant neoplasm of ovary; Mediating; Messenger RNA; Mitotic; Mitotic spindle; Modification; Neurodegenerative Disorders; Organism; Oxidative Stress; Pathway interactions; Pharmacologic Substance; Phase III Clinical Trials; Play; Poly Adenosine Diphosphate Ribose; Poly(ADP-ribose) Polymerases; Polymerase; Polymers; Post-Transcriptional Regulation; Post-Translational Protein Processing; Process; Property; Protein Binding; Protein Family; Proteins; RNA; RNA Binding; RNA Recognition Motif; RNA Viruses; RNA-Binding Proteins; RRM1 gene; RRM2 gene; Recruitment Activity; Regulation; Ribosomes; Role; Set protein; Site; Solid Neoplasm; Specificity; Stress; Structure; Structure-Activity Relationship; Testing; Time; Transcriptional Regulation; Translations; Virus Diseases; Work; Zinc Fingers; biological adaptation to stress; cancer therapy; effective therapy; genetic regulatory protein; human disease; inhibitor/antagonist; insight; mRNA Stability; mRNA Transcript Degradation; malignant breast neoplasm; member; monomer; novel; overexpression; protein complex; research study; response; scaffold

DESCRIPTION (provided by applicant): Poly(ADP-ribose) (pADPr) is a unique polymer required for life in multicellular organisms. It functions as both a covalent modification of acceptor proteins and as a scaffold that binds specific sets of proteins. The polymer plays a role in essential cellular functions including cell division and cell cycle progression and regulation of transcription and translation. pADPr also functions in cell stress responses such as apoptosis, DNA damage repair, and innate immune responses. In preliminary results we identified a new function for pADPr and the enzymes that polymerize it, pADPr polymerases (PARPs) in regulation of stress granule (SG) assembly and the post-transcriptional regulation of mRNAs. These results connect the fields of cellular stress, PARPs and mRNA regulation in an unexpected manner. They suggest that pADPr functions as a structural scaffold for SG assembly, similar to its function at the spindle pole and at sites of DNA damage. We anticipate that our results will have a high impact on all three fields and wish to extend our work to include mechanistic studies. One of our long-term goals is to understand how pADPr functions as a scaffold. SG assembly and mRNA regulation is an ideal process to study the scaffold function of pADPr. In this proposal, we seek to determine the mechanism of pADPr function in the assembly of a SG and begin to understand how pADPr binding to proteins regulates function. We do so using a combination of biochemical assays and cell biology. In specific Aim 1 we identify the sites of pADPr modification that are required for SG assembly and generate and determine the mechanism in which pADPr is synthesized for stress. In Aim 2 we determine the manner in which pADPr regulates mRNA binding and recruitment to the SG, and in Aim 3 we examine the structure- function relationships that govern binding of pADPr to proteins. We believe the proposed experiments will help elucidate the scaffold function of pADPr elsewhere in the cell. SGs have important disease relevance. PARP inhibition, already shown to be effective for breast and ovarian cancer therapies, might be equally effective for treatment of other stress-related diseases such as solid tumors, ischemia, and neurodegenerative disease.

描述（由申请人提供）：poly（adp-ribose）（PADPR）是多细胞生物生命所需的独特聚合物。它既是对受体蛋白的共价修饰，又是结合特定蛋白质集的支架。聚合物在必需的细胞功能中起作用，包括细胞分裂和细胞周期进程以及转录和翻译的调节。 PADPR还可以在细胞应激反应中起作用，例如凋亡，DNA损伤修复和先天免疫反应。在初步结果中，我们确定了PADPR的新功能和将其聚合的酶，PADPR聚合酶（PARPS（PARP）调节，以调节应力颗粒（SG）组装（SG）组装和mRNA的转录后调节。这些结果以意想不到的方式连接细胞应力，PARP和mRNA调节的场。他们认为PADPR是SG组装的结构支架，类似于其在纺锤极和DNA损伤部位的功能。我们预计我们的结果将对这三个领域产生很大的影响，并希望将我们的工作扩展到包括机械研究。 我们的长期目标之一是了解PADPR如何充当脚手架。 SG组装和mRNA调控是研究PADPR的支架函数的理想过程。在此提案中，我们试图确定SG组装中PADPR功能的机理，并开始了解PADPR与蛋白质的结合如何调节功能。我们使用生化测定和细胞生物学的组合来做到这一点。在特定的目标1中，我们确定了SG组装所需的PADPR修改位点，并生成并确定合成PADPR的应力机制。在目标2中，我们确定了PADPR调节mRNA结合和募集与SG的方式，在AIM 3中，我们检查了控制PADPR与蛋白质结合的结构 - 功能关系。我们认为，提出的实验将有助于阐明细胞其他地方PADPR的支架功能。 SG具有重要的疾病相关性。 PARP抑制作用已经证明对乳腺癌和卵巢癌疗法有效，可能同样有效地治疗其他与压力有关的疾病，例如实体瘤，缺血和神经退行性疾病。